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Highlights • Effect of droplet banding on demulsification process is analyzed. • Optimal acoustic intensity and treatment time are obtained. • Acoustic cavitation threshold in emulsions is achieved by spectral analysis. • Range of oil-water properties suitable for ultrasonic separation is determined. Abstract This paper focuses on the application range and optimal parameters for separating water-in-oil (W/O) emulsions using ultrasonic standing waves (USWs). Firstly, the effect of droplet bandings on the demulsification process is theoretically analyzed. The formation of droplet bandings coincides with droplet sedimentation when horizontal irradiation is applied, which improves the separation efficiency. On this basis, the effects of acoustic parameters and oil-water properties on demulsification are analyzed in detail. It is shown that ultrasonic treatment is an efficient demulsification method, and that maximum demulsification efficiency can be achieved by optimizing the acoustic intensity and treatment time. Excessive acoustic intensity causes acoustic cavitation in low-frequency USWs and acoustic streaming in high-frequency USWs, resulting in a low separation efficiency. Acoustic cavitation also causes secondary emulsification, dispersing tiny oil droplets into separated water. In addition, ultrasonic separation is significantly affected by oil-water properties. The demulsification efficiency of W/O emulsions decreases with increasing oil viscosity and decreasing oil-water interfacial tension. Ultrasonic treatment has significant advantages for separation of emulsions with low water content and interfacial tension. This study presents an in-depth analysis of ultrasonic separation. [ABSTRACT FROM AUTHOR]